Prognostic impact of FOXP3 expression in triple-negative breast cancer

Figures & data

Figure 1. Patients’ cohort.

Table I. Correlation analyses between the number of FOXP3-positive Tregs and both clinicopathologic data for 86 TNBC patients.

		Regulatory T cells
Variable Total number of patients, n (%)		FOXP3+ < 15 64 (74.4%)	FOXP3+ ≥ 15 22 (25.6%)	p-value*
Age, years	< 45	36 (56.3%)	7 (31.8%)	0.085
	≥ 45	28 (44.0%)	15 (68.0%)
Histologic grade	Low-intermediate	13 (20.5%)	1 (4.5%)	0.052
	High	39 (60.8%)	21 (95.5%)
	Missing	12 (18.7%)	0 (0.0%)
Initial stage	l	10 (15.6%)	3 (13.6%)	0.744
	ll	26 (40.6%)	11 (50.0%)
	lll	28 (43.8%)	8 (36.4%)
Lymphovascular invasion	Yes	20 (31.3%)	13 (59.1%)	0.032
	No	17 (26.6%)	7 (31.8%)
	Missing	27 (42.1%)	2 (3.1%)
Extensive intraductal component	Yes	4 (6.3%)	3 (13.6%)	1.000
	No	32 (50.0%)	19 (86.4%)
	Missing	28 (43.7%)	0 (0.0%)
Adj chemotherapy	Yes	58 (90.6%)	22 (100.0%)	0.331
	No	6 (9.4%)	0 (0.0%)
Adj anthra	Yes	45 (70.3%)	18 (81.8%)	0.405
	No	19 (29.7%)	4 (18.2%)
Adj taxane	Yes	16 (25.0%)	11 (50.0%)	0.048
	No	48 (75.0%)	11 (50.0%)
Metastatic sites metastatic LNs	Yes	24 (37.5%)	14 (63.6%)	0.048
	No	40 (62.5%)	8 (36.4%)
bone	Yes	23 (35.9%)	8 (36.4%)	1.000
	No	41 (64.1%)	14 (63.6%)
liver	Yes	13 (20.3%)	2 (9.0%)	0.333
	No	51 (79.7%)	20 (91.0%)
lung	Yes	32 (50.0%)	8 (36.4%)	0.326
	No	32 (50.0%)	14 (63.6%)
pleura	Yes	15 (23.4%)	0 (0.0%)	0.008
	No	49 (76.6%)	22 (100.0%)
brain	Yes	8 (12.5%)	4 (18.2%)	0.504
	No	56 (87.5%)	18 (81.8%)
Relapse free survival (months)		20.5 (17.8 – 23.2)	20.3 (11.9 – 28.7)	0.495
Progression free survival (months)		3.0 (1.9 – 4.1)	4.2 (0.6 – 7.8)	0.036
Overall survival (months)		16.8 (12.2 – 23.4)	23.8 (13.8 – 88.2)	0.001

Adj, adjuvant; Anthra, anthracycline; FOXP3, forkhead box protein P3; LN, lymph node; Tregs, regulatory T cells;

*The χ²-test was used to test for differences between categorical variables.

Figure 2. FOXP3 immunohisochemical staining. Intracelluar nuclear stainings (black arrows) indicate positive FOXP3 lymphocytes in peritumoral area underling invasive ductal carcinoma background (× 400).

Table II. Univariate analysis for metastatic overall survival and progression free survival to first line palliative chemotherapy in the 86 patients.

Variables	No.	OS		PFS
		Months (95% CI)	p-value	Months (95% CI)	p-value
Age ≥ 45
No	43	17.2 (7.6 − 26.8)	0.235	3.7 (3.2 − 4.2)	0.806
Yes	43	21.5 (11.3 − 31.7)		3.5 (1.6 − 5.4)
HG lll (n = 74)
No	14	16.7	0.371	4.3 (3.3 − 5.3)	0.469
Yes	60	18.5 (15.4 − 21.6)		3.7 (2.8 − 4.6)
Stage lll
No	49	21.5 (10.4 − 32.6)	0.116	4.1 (3.2 − 4.7)	0.076
Yes	37	13.4 (5.2 − 21.6)		2.5 (1.8 − 3.5)
LVI (n = 57)
No	24	17.2 (6.6 − 27.8)	0.402	3.0 (0.5 − 5.5)	0.058
Yes	33	24.3 (17.8 − 31.8)		4.0 (2.8 − 5.2)
EIC (n = 68)
No	51	18.5 (14.0 − 23.0)	0.552	3.8 (2.6 − 5.0)	0.441
Yes	7	50.9 (19.0 − 82.8)		11.0 (0.0 − 31.5)
Adj Anthra
No	23	12.5 (9.2 − 15.8)	0.132	3.9 (3.3 − 4.5)	0.896
Yes	63	21.2 (17.0 − 25.4)		3.5 (2.4 − 4.6)
Adj Taxane
No	59	20.1 (12.5 − 27.7)	0.650	3.6 (2.2 − 5.0)	0.921
Yes	27	18.5 (12.0 − 25.0)		3.8 (2.5 − 5.2)
FOXP3+
< 15	64	16.8 (12.2 − 23.4)	0.001	3.0 (1.9 − 4.1)	0.036
≥ 15	22	23.8 (13.8 − 88.2)		4.2 (0.6 − 7.8)

Adj, adjuvant; Anthra, anthracycline; CI, confidence interval; EIC, extensive intraductal component; FOXP3, forkhead box protein P3; HG, histologic grade; LVI, lymphovasculari invasion; No., number; OS, overall survival; PFS, progression free survival.

Figure 3. Kaplan-Meier Survival curves for PFS and OS. (A) showed PFS curves according to the status of positive FOXP3 cells. Analysis of PFS revealed a significantly better prognosis for FOXP3+ Tregs ≥ 15 compared with FOXP3+ Tregs < 15 (p = 0.036). (B) showed OS curves according to the status of positive FOXP3 cells. Analysis of OS revealed a significantly better prognosis for FOXP3+ Tregs ≥ 15 compared with FOXP3+ Tregs < 15 (p = 0.001).

Table III. Multivariate proportional analysis of overall survival in TNBC patients.

Variable	OS			PFS
	Hazard ratio	95% CI	p-value	Hazard ratio	95% CI	p-value
FOXP3 Treg < 15	2.4	1.0 − 5.6	0.049	2.0	1.1 − 3.6	0.032
Stage lll				0.3	0.1 − 0.6	0.001
LVI +				3.2	1.5 − 6.5	0.002

CI, confidence interval; FOXP3, forkhead box protein P3; LVI, lymphovascular invasion.

Table IV. Clinicopathologic characteristics of the patients who had 48 months or longer metastatic overall survival.

Pts	No. of FOXP3	Age	Stage	Adj CTx	Histology	HG	RFS (ms)	Initial metastatic sites	Response to first line palliative chemotherapy	PFS (ms)	# of palliative chemotherapy lines	mOS (ms)
1	33	49	IIA	Yes	IDC	High	31.3	Bone & LNs	PR	7.0	3	48.4
2	17	49	IIA	Yes	IDC	High	41.2	Bone	SD	21.0	3	52.9+
3	34	47	IIIA	Yes	Metaplastic	High	18.3	Lung	PD	1.9	4	53.2+
4	72	38	IIIC	Yes	IDC	High	54.8	LNs	CR	28.0	4	60.3+
5	66	45	IIA	Yes	IDC	High	35.9	Bone & LNs	PR	13.4	4	60.8
6	161	52	I	Yes	IDC	High	19.2	Lung & LNs	CR	11.0	5	60.9+
7	24	36	IIA	Yes	IDC	Intermediate	73.3	Lung	SD	4.7	5	61.2
8	18	51	I	Yes	IDC	High	56.9	Lung	SD	4.0	6	70.7+
9	40	28	IIIA	Yes	IDC	High	34.8	Lung	PR	47.0	6	72.7
10	66	32	IIIC	Yes	IDC	High	80.3	LNs	PR	12.0	7	78.1+
11	27	54	I	Yes	IDC	High	8.9	Bone	SD	12.5	5	88.2+

Adj CTx, adjuvant chemotherapy; CR, complete response; HG, histologic grade; IDC, invasive ductal carcinoma; Interm, intermediate; mOS, metastatic overall survival; ms, months; No., number; PD, progressive disease; +, alive at present; PFS, progression-free survival; PR, partial response; Pts, Patients; RFS, relapse-free survival; SD, stable disease; stage, initial stage.